Mapping of the 5′ terminal nucleotides of Grapevine leafroll-associated virus 3 sgRNAs

Grapevine leafroll-associated virus 3 (GLRaV-3) is a positive sense, single stranded RNA virus that has a detrimental effect on wine and table grapes worldwide. Previous studies have shown that GLRaV-3, like other closteroviruses produces subgenomic (sg) RNAs during replication and that these sgRNAs are deployed for the expression of the ORFs on the 3′ half of the genome. In this study a dsRNA blot confirmed the presence of three, 3′ co-terminal sgRNAs species [sgRNA(ORF3/4), sgRNA(ORF5) and sgRNA(ORF6)] in GLRaV-3-infected plant material when using a probe directed at the coat protein. The specific 5′ terminal nucleotides for these sgRNAs as well as four additional sgRNAs [sgRNA(ORF7), sgRNA(ORF8), sgRNA(ORF9) and sgRNA(ORF10-12)] were determined by RLM-RACE for GLRaV-3 isolate GP18.